title: Purification and characterization of a novel thermostable luciferase from Benthosema pterotum.
abstract: A novel luciferase from Benthosema pterotum, collected from Port of Jask, close to Persian Gulf, was purified for the first time, using Q-Sepharose anion exchange chromatography. The molecular mass of the novel enzyme, measured by SDS-PAGE technique, was about 27 kDa and its Km value is 0.4 μM; both values are similar to those of other coelenterazine luciferases. B. pterotum (BP) luciferase showed maximum intensity of emitted light at 40°C, in 20mM Tris buffer, pH 9 and 20 mM magnesium concentration. Experimental measurements indicated that BP luciferase is a relatively thermostable enzyme; furthermore it shows a high residual activity at extreme pH values. Its biological activity is strongly inhibited by 1 mM Cu(2+), Zn(2+) and Ni(2+), while calcium and mainly magnesium ions strongly increase BP luciferase activity. The B. pterotum luciferase generated blue light with a maximum emission wavelength at 475 nm and showed some similarity with other luciferases, while other parameters appeared quite different, in this way, confirming that a novel protein has been purified.
authors.affiliation: Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Ahmad Abolpour
authors.initials: AA
authors.lastname: Homaei
authors.forename: Asma Bahari
authors.initials: AB
authors.lastname: Mymandi
authors.forename: Reyhaneh
authors.initials: R
authors.lastname: Sariri
authors.forename: Ehsan
authors.initials: E
authors.lastname: Kamrani
authors.forename: Roberto
authors.initials: R
authors.lastname: Stevanato
authors.forename: Seyed-Masoud
authors.initials: SM
authors.lastname: Etezad
authors.forename: Khosro
authors.initials: K
authors.lastname: Khajeh
mesh_terms: D000818:Animals; D005399:Fishes / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008156:Luciferases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D049449:Luminescence; D008670:Metals / Q000737:chemistry
chemical_list: D008670:Metals; D008156:Luciferases
keywords: Benthosema pterotum luciferase; Bioluminescence; Enzyme purification; Persian Gulf